Natural human and murine IL-6 are glycoproteins containing N-and/or O-linked carbohydrates (human IL-6 contains two potential N-glycosylation sites, while mouse IL-6 has none). In comparison with mouse IL-6, human IL-6 exhibits approximately 65% sequence homology at the nucleotide level, and 42% homology at the amino acid level. Although human and mouse IL-6 are equally active on mouse cells, mouse IL-6 is not active on human cells.

IL-6 is predicted to have a four helix-bundle type tertiary structure found in a number of other cytokines including growth hormone, EPO, G-CSF, OSM, IL-11, CNTF, LIF, MGF, prolactin, etc. The gene structures of these cytokines also show varying degrees of relatedness. Based on these criteria, it has been suggested that these cytokines evolved from a common ancestral gene. Results of structure-function studies of IL-6 and other four a-helix bundle cytokines indicated that the c-terminal (helix D) regions of these cytokines are primarily responsible for binding to the receptors.

Interleukin 6 exerts multiple functions on numerous target cells. IL-6 plays an important role in immune functions. It has effects on B cell differentiation and antibody production, on cytotoxic T cell differentiation, on T cell activation, growth and differentiation, and on the induction of IL-2Ra chain expression and IL-2 production in T cells. In hemopoiesis, IL-6 has blast cell growth factor activity and can synergize with IL-3 to shorten the G0 period of early hemopoietic progenitors. In addition, IL-6 has been found to synergize with IL-3 in megakaryocyte development, increasing platelet numbers in vivo and the number, size and average ploidy value of megakaryocyte colonies formed from mouse or human bone marrow cells in vitro. Similarly to IL-11 and LIF, IL-6 can induce the synthesis of hepatic acute phase proteins both in vivo and in vitro. IL-6 has growth factor activities and will stimulate the growth of hybridomas, plasmacytomas, myelomas, sarcomas, carcinomas, EBV-transformed B cells, keratinocytes, and mesangial cells. In contrast to its growth stimulatory activities, IL-6 is also a growth inhibitor for a number of leukemia and carcinoma cell lines. Additional bioactivities attributed to IL-6 include, induction of terminal differentiation of M1 myeloid leukemic cells; the differentiation and survival of neuronal cells; and the activation of osteoclast development. Although IL-6 was also discovered as an antiviral factor produced by human diploid fibroblasts, the question of whether or not IL-6 has antiviral activity is controversial. Many groups have been consistently unable to find any antiviral activity for recombinant human IL-6. The various activities of IL-6 described above suggest that this factor will have a major role in the mediation of the inflammatory and immune responses initiated by infection or injury. Although the exact functions of IL-6 in vivo are not known, elevated IL-6 levels have been reported to be associated with a variety of diseases, including autoimmune diseases, mesangial proliferative glomerulonephritis, psoriasis, and malignancies such as plasmacytoma and myeloma. A great deal of work is currently in progress in order to provide a better understanding of the role of IL-6 in the modulation of normal and pathological processes. This IL-6 ELISA is a 3.5 hour solid phase immunoassay readily applicable to measure IL-6 levels in serum, plasma, cell culture supernatant, and other biological fluids in the range of 12.5-400pg/ml. It showed no cross reactivity with various other cytokines superfamily proteins.

Calibrator Diluent I: 2.0pg/ml Calibrator Diluent II: 1.75pg/ml

12.5-400pg/ml

This sandwich ELISA recognizes both natural and recombinant human IL-6, exhibiting no detectable crossreactivity with human; SAA, EGF, IL-8, IL-16, MCAF, MCP-3, TNF-g, M-CSF, GM-CSF, TGF-ß1, RANTES, FGF, MIP-1a, CRP, EPO, IFN-g.

I8428-04A: IL-6 Microtiter Plate 1x96 wells, Pre-coated with anti-human IL-6 monoclonal antibody. I8428-04B: IL-6 Mab (Biotin) 1x6ml, Anti-human IL-6 monoclonal antibody conjugated to Biotin. I8428-04C: Avidin (HRP) 1x12ml, Avidin conjugated to horseradish peroxidase. *I8428-04D: IL-6 Standard 2x1 vial, Recombinant human IL-6 (800pg/vial) in a buffered protein base with preservative, lyophilized. I8428-04E: Calibrator Diluent I 1x25ml, Animal serum with buffer and preservative. For serum/plasma testing. I8428-04F: Calibrator Diluent II, 5X 1x25ml, Cell culture medium with calf serum and preservative. For cell culture supernatant testing. I8428-04G: Wash Buffer, 20X 1x60ml, 20-fold concentrated solution of buffered surfactant. I8428-04H: Substrate A, 1x10ml, Buffered solution with H2O2 I8428-04J: Substrate B, 1x10ml, Buffered solution with TMB. I8428-04K: Stop Solution, 1x14ml, 2N Sulfuric Acid (H2SO4)

Store *I8428-04D powder at 4°C liquid at -20°C (30 days). Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.